Binding of GDNF and neurturin to human GDNF family receptor alpha 1 and 2. Influence of cRET and cooperative interactions
| Autor: | M, Cik, S, Masure, A S, Lesage, I, Van Der Linden, P, Van Gompel, M N, Pangalos, R D, Gordon, J E, Leysen |
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| Rok vydání: | 2000 |
| Předmět: |
Glial Cell Line-Derived Neurotrophic Factor Receptors
Recombinant Fusion Proteins Neurturin Proto-Oncogene Proteins c-ret Receptor Protein-Tyrosine Kinases Nerve Tissue Proteins Receptor Cross-Talk Iodine Radioisotopes Substantia Nigra Kinetics Radioligand Assay Proto-Oncogene Proteins Drosophila Proteins Humans Protein Isoforms Glial Cell Line-Derived Neurotrophic Factor Nerve Growth Factors Cloning Molecular Gene Library |
| Zdroj: | The Journal of biological chemistry. 275(36) |
| ISSN: | 0021-9258 |
| Popis: | The members of the glial cell line-derived neurotrophic factor (GDNF) family signal via binding to the glycosyl phosphatidylinositol-anchored membrane proteins, the GDNF family receptors alpha (GFRalpha), and activation of cRET. We performed a detailed analysis of the binding of GDNF and neurturin to their receptors and investigated the influence of cRET on the binding affinities. We show that the rate of dissociation of (125)I-GDNF from GFRalpha1 is increased in the presence of 50 nm GDNF, an effect that can be explained by the occurrence of negative cooperativity. Scatchard plots of the ligand concentration binding isotherms reveal a pronounced downward curvature at low (125)I-GDNF concentrations suggesting the presence of positive cooperativity. This effect is observed in the range of GDNF concentrations responsible for biological activity (1-20 pm) and may have an important role in cRET-independent signaling. A high affinity site with a K(D) of 11 pm for (125)I-GDNF is detected only when GFRalpha1 is co-expressed with cRET at a DNA ratio of 1:3. These results suggest an interaction of GFRalpha1 and cRET in the absence of GDNF and demonstrate that the high affinity binding can be measured only when cRET is present. |
| Databáze: | OpenAIRE |
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